                         SEQUENCE LISTING

<110>  GE HEALTHCARE BIO-SCIENCES CORP.
       MITSIS, Paul
       SEKHER, Anuradha
       KUMAR, Gyanendra
 
<120>  A SINGLE ENZYME SYSTEM FOR FAST, ULTRA LONG PCR

<130>  PB07115

<150>  US 61/016,850
<151>  2007-12-27

<150>  US 61/025,047
<151>  2008-01-31

<160>  39    

<170>  PatentIn version 3.3

<210>  1
<211>  778
<212>  PRT
<213>  Thermococcus barossii

<400>  1

Met Ile Leu Asp Val Asp Tyr Ile Thr Glu Asp Gly Lys Pro Val Ile 
1               5                   10                  15      


Arg Val Phe Lys Lys Asp Lys Gly Glu Phe Lys Ile Glu Tyr Asp Arg 
            20                  25                  30          


Glu Phe Glu Pro Tyr Ile Tyr Ala Leu Leu Arg Asp Asp Ser Ala Ile 
        35                  40                  45              


Glu Glu Ile Glu Lys Ile Thr Ala Glu Arg His Gly Lys Val Val Lys 
    50                  55                  60                  


Val Lys Arg Ala Glu Lys Val Lys Lys Lys Phe Leu Gly Arg Ser Val 
65                  70                  75                  80  


Glu Val Trp Val Leu Tyr Phe Thr His Pro Gln Asp Val Pro Ala Ile 
                85                  90                  95      


Arg Asp Lys Ile Arg Lys His Pro Ala Val Ile Asp Ile Tyr Glu Tyr 
            100                 105                 110         


Asp Ile Pro Phe Ala Lys Arg Tyr Leu Ile Asp Lys Gly Leu Val Pro 
        115                 120                 125             


Met Glu Gly Asp Glu Glu Leu Lys Leu Met Ser Phe Asp Ile Glu Thr 
    130                 135                 140                 


Leu Tyr His Glu Gly Glu Glu Phe Gly Thr Gly Pro Ile Leu Met Ile 
145                 150                 155                 160 


Ser Tyr Ala Asp Glu Ser Glu Ala Arg Val Ile Thr Trp Lys Lys Ile 
                165                 170                 175     


Asp Leu Pro Tyr Val Asp Val Val Ser Thr Glu Lys Glu Met Ile Lys 
            180                 185                 190         


Arg Phe Leu Lys Val Val Lys Glu Lys Asp Pro Asp Val Leu Ile Thr 
        195                 200                 205             


Tyr Asn Gly Asp Asn Phe Asp Phe Ala Tyr Leu Lys Lys Arg Cys Glu 
    210                 215                 220                 


Lys Leu Gly Val Ser Phe Thr Leu Gly Arg Asp Gly Ser Glu Pro Lys 
225                 230                 235                 240 


Ile Gln Arg Met Gly Asp Arg Phe Ala Val Glu Val Lys Gly Arg Ile 
                245                 250                 255     


His Phe Asp Leu Tyr Pro Val Ile Arg Arg Thr Ile Asn Leu Pro Thr 
            260                 265                 270         


Tyr Thr Leu Glu Ala Val Tyr Glu Ala Val Phe Gly Lys Pro Lys Glu 
        275                 280                 285             


Lys Val Tyr Ala Glu Glu Ile Ala Thr Ala Trp Glu Thr Gly Glu Gly 
    290                 295                 300                 


Leu Glu Arg Val Ala Arg Tyr Ser Met Glu Asp Ala Arg Val Thr Tyr 
305                 310                 315                 320 


Glu Leu Gly Arg Glu Phe Phe Pro Met Glu Ala Gln Leu Ser Arg Leu 
                325                 330                 335     


Ile Gly Gln Gly Leu Trp Asp Val Ser Arg Ser Ser Thr Gly Asn Leu 
            340                 345                 350         


Val Glu Trp Phe Leu Leu Arg Lys Ala Tyr Glu Arg Asn Glu Leu Ala 
        355                 360                 365             


Pro Asn Lys Pro Asp Glu Arg Glu Leu Ala Arg Arg Arg Gly Gly Tyr 
    370                 375                 380                 


Ala Gly Gly Tyr Val Lys Glu Pro Glu Arg Gly Leu Trp Asp Asn Ile 
385                 390                 395                 400 


Val Tyr Leu Asp Phe Arg Ser Leu Tyr Pro Ser Ile Ile Ile Thr His 
                405                 410                 415     


Asn Val Ser Pro Asp Thr Leu Asn Arg Glu Gly Cys Lys Ser Tyr Asp 
            420                 425                 430         


Val Ala Pro Gln Val Gly His Lys Phe Cys Lys Asp Phe Pro Gly Phe 
        435                 440                 445             


Ile Pro Ser Leu Leu Gly Asn Leu Leu Glu Glu Arg Gln Lys Ile Lys 
    450                 455                 460                 


Arg Lys Met Lys Ala Thr Leu Asp Pro Leu Glu Arg Lys Leu Leu Asp 
465                 470                 475                 480 


Tyr Arg Gln Arg Ala Ile Lys Ile Leu Ala Asn Ser Phe Tyr Gly Tyr 
                485                 490                 495     


Tyr Gly Tyr Ala Arg Ala Arg Trp Tyr Cys Lys Glu Cys Ala Glu Ser 
            500                 505                 510         


Val Thr Ala Trp Gly Arg Glu Tyr Ile Glu Met Val Ile Arg Glu Leu 
        515                 520                 525             


Glu Glu Lys Phe Gly Phe Lys Val Leu Tyr Ala Asp Thr Asp Gly Leu 
    530                 535                 540                 


His Ala Thr Ile Pro Gly Ala Asp Ala Glu Thr Val Lys Lys Lys Ala 
545                 550                 555                 560 


Met Glu Phe Leu Asn Tyr Ile Asn Pro Lys Leu Pro Gly Leu Leu Glu 
                565                 570                 575     


Leu Glu Tyr Glu Gly Phe Tyr Val Arg Gly Phe Phe Val Thr Lys Lys 
            580                 585                 590         


Lys Tyr Ala Val Ile Asp Glu Glu Gly Lys Ile Thr Thr Arg Gly Leu 
        595                 600                 605             


Glu Ile Val Arg Arg Asp Trp Ser Glu Ile Ala Lys Glu Thr Gln Ala 
    610                 615                 620                 


Arg Val Leu Glu Ala Ile Leu Arg His Gly Asp Val Glu Glu Ala Val 
625                 630                 635                 640 


Arg Ile Val Lys Glu Val Thr Glu Lys Leu Ser Lys Tyr Glu Val Pro 
                645                 650                 655     


Pro Glu Lys Leu Val Ile His Glu Gln Ile Thr Arg Glu Leu Lys Asp 
            660                 665                 670         


Tyr Lys Ala Thr Gly Pro His Val Ala Ile Ala Lys Arg Leu Ala Ala 
        675                 680                 685             


Arg Gly Ile Lys Ile Arg Pro Gly Thr Val Ile Ser Tyr Ile Val Leu 
    690                 695                 700                 


Lys Gly Ser Gly Arg Ile Gly Asp Arg Ala Ile Pro Phe Asp Glu Phe 
705                 710                 715                 720 


Asp Pro Thr Lys His Arg Tyr Asp Ala Asp Tyr Tyr Ile Glu Asn Gln 
                725                 730                 735     


Val Leu Pro Ala Val Glu Arg Ile Leu Arg Ala Phe Gly Tyr Lys Lys 
            740                 745                 750         


Glu Asp Leu Arg Tyr Gln Lys Thr Arg Gln Val Gly Leu Gly Ala Trp 
        755                 760                 765             


Leu Gly Met Gly Gly Glu Arg Leu Lys Leu 
    770                 775             


<210>  2
<211>  778
<212>  PRT
<213>  Thermococcus barossii

<400>  2

Met Ile Leu Asp Val Asp Tyr Ile Thr Glu Asp Gly Lys Pro Val Ile 
1               5                   10                  15      


Arg Val Phe Lys Lys Asp Lys Gly Glu Phe Lys Ile Glu Tyr Asp Arg 
            20                  25                  30          


Glu Phe Glu Pro Tyr Ile Tyr Ala Leu Leu Arg Asp Asp Ser Ala Ile 
        35                  40                  45              


Glu Glu Ile Glu Lys Ile Thr Ala Glu Arg His Gly Lys Val Val Lys 
    50                  55                  60                  


Val Lys Arg Ala Glu Lys Val Lys Lys Lys Phe Leu Gly Arg Ser Val 
65                  70                  75                  80  


Glu Val Trp Val Leu Tyr Phe Thr His Pro Gln Asp Val Pro Ala Ile 
                85                  90                  95      


Arg Asp Lys Ile Arg Lys His Pro Ala Val Ile Asp Ile Tyr Glu Tyr 
            100                 105                 110         


Asp Ile Pro Phe Ala Lys Arg Tyr Leu Ile Asp Lys Gly Leu Val Pro 
        115                 120                 125             


Met Glu Gly Asp Glu Glu Leu Lys Leu Met Ser Phe Ala Ile Ala Thr 
    130                 135                 140                 


Leu Tyr His Glu Gly Glu Glu Phe Gly Thr Gly Pro Ile Leu Met Ile 
145                 150                 155                 160 


Ser Tyr Ala Asp Glu Ser Glu Ala Arg Val Ile Thr Trp Lys Lys Ile 
                165                 170                 175     


Asp Leu Pro Tyr Val Asp Val Val Ser Thr Glu Lys Glu Met Ile Lys 
            180                 185                 190         


Arg Phe Leu Lys Val Val Lys Glu Lys Asp Pro Asp Val Leu Ile Thr 
        195                 200                 205             


Tyr Asn Gly Asp Asn Phe Asp Phe Ala Tyr Leu Lys Lys Arg Cys Glu 
    210                 215                 220                 


Lys Leu Gly Val Ser Phe Thr Leu Gly Arg Asp Gly Ser Glu Pro Lys 
225                 230                 235                 240 


Ile Gln Arg Met Gly Asp Arg Phe Ala Val Glu Val Lys Gly Arg Ile 
                245                 250                 255     


His Phe Asp Leu Tyr Pro Val Ile Arg Arg Thr Ile Asn Leu Pro Thr 
            260                 265                 270         


Tyr Thr Leu Glu Ala Val Tyr Glu Ala Val Phe Gly Lys Pro Lys Glu 
        275                 280                 285             


Lys Val Tyr Ala Glu Glu Ile Ala Thr Ala Trp Glu Thr Gly Glu Gly 
    290                 295                 300                 


Leu Glu Arg Val Ala Arg Tyr Ser Met Glu Asp Ala Arg Val Thr Tyr 
305                 310                 315                 320 


Glu Leu Gly Arg Glu Phe Phe Pro Met Glu Ala Gln Leu Ser Arg Leu 
                325                 330                 335     


Ile Gly Gln Gly Leu Trp Asp Val Ser Arg Ser Ser Thr Gly Asn Leu 
            340                 345                 350         


Val Glu Trp Phe Leu Leu Arg Lys Ala Tyr Glu Arg Asn Glu Leu Ala 
        355                 360                 365             


Pro Asn Lys Pro Asp Glu Arg Glu Leu Ala Arg Arg Arg Gly Gly Tyr 
    370                 375                 380                 


Ala Gly Gly Tyr Val Lys Glu Pro Glu Arg Gly Leu Trp Asp Asn Ile 
385                 390                 395                 400 


Val Tyr Leu Asp Phe Arg Ser Leu Tyr Pro Ser Ile Ile Ile Thr His 
                405                 410                 415     


Asn Val Ser Pro Asp Thr Leu Asn Arg Glu Gly Cys Lys Ser Tyr Asp 
            420                 425                 430         


Val Ala Pro Gln Val Gly His Lys Phe Cys Lys Asp Phe Pro Gly Phe 
        435                 440                 445             


Ile Pro Ser Leu Leu Gly Asn Leu Leu Glu Glu Arg Gln Lys Ile Lys 
    450                 455                 460                 


Arg Lys Met Lys Ala Thr Leu Asp Pro Leu Glu Arg Lys Leu Leu Asp 
465                 470                 475                 480 


Tyr Arg Gln Arg Ala Ile Lys Ile Leu Ala Asn Ser Phe Tyr Gly Tyr 
                485                 490                 495     


Tyr Gly Tyr Ala Arg Ala Arg Trp Tyr Cys Lys Glu Cys Ala Glu Ser 
            500                 505                 510         


Val Thr Ala Trp Gly Arg Glu Tyr Ile Glu Met Val Ile Arg Glu Leu 
        515                 520                 525             


Glu Glu Lys Phe Gly Phe Lys Val Leu Tyr Ala Asp Thr Asp Gly Leu 
    530                 535                 540                 


His Ala Thr Ile Pro Gly Ala Asp Ala Glu Thr Val Lys Lys Lys Ala 
545                 550                 555                 560 


Met Glu Phe Leu Asn Tyr Ile Asn Pro Lys Leu Pro Gly Leu Leu Glu 
                565                 570                 575     


Leu Glu Tyr Glu Gly Phe Tyr Val Arg Gly Phe Phe Val Thr Lys Lys 
            580                 585                 590         


Lys Tyr Ala Val Ile Asp Glu Glu Gly Lys Ile Thr Thr Arg Gly Leu 
        595                 600                 605             


Glu Ile Val Arg Arg Asp Trp Ser Glu Ile Ala Lys Glu Thr Gln Ala 
    610                 615                 620                 


Arg Val Leu Glu Ala Ile Leu Arg His Gly Asp Val Glu Glu Ala Val 
625                 630                 635                 640 


Arg Ile Val Lys Glu Val Thr Glu Lys Leu Ser Lys Tyr Glu Val Pro 
                645                 650                 655     


Pro Glu Lys Leu Val Ile His Glu Gln Ile Thr Arg Glu Leu Lys Asp 
            660                 665                 670         


Tyr Lys Ala Thr Gly Pro His Val Ala Ile Ala Lys Arg Leu Ala Ala 
        675                 680                 685             


Arg Gly Ile Lys Ile Arg Pro Gly Thr Val Ile Ser Tyr Ile Val Leu 
    690                 695                 700                 


Lys Gly Ser Gly Arg Ile Gly Asp Arg Ala Ile Pro Phe Asp Glu Phe 
705                 710                 715                 720 


Asp Pro Thr Lys His Arg Tyr Asp Ala Asp Tyr Tyr Ile Glu Asn Gln 
                725                 730                 735     


Val Leu Pro Ala Val Glu Arg Ile Leu Arg Ala Phe Gly Tyr Lys Lys 
            740                 745                 750         


Glu Asp Leu Arg Tyr Gln Lys Thr Arg Gln Val Gly Leu Gly Ala Trp 
        755                 760                 765             


Leu Gly Met Gly Gly Glu Arg Leu Lys Leu 
    770                 775             


<210>  3
<211>  20
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  3
gctgaagtgg tggaaaccgc                                                 20


<210>  4
<211>  22
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  4
acagccaagc ttgcagaaac ga                                              22


<210>  5
<211>  22
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  5
aacgtgtccg cgcctttgat tt                                              22


<210>  6
<211>  24
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  6
tttcctgaca gtgacagact gcgt                                            24


<210>  7
<211>  21
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  7
gcctcgcata tcaggaagca c                                               21


<210>  8
<211>  20
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  8
cgcgcggtat tgctaacacg                                                 20


<210>  9
<211>  20
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  9
gattcgccag acgattgaac                                                 20


<210>  10
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  10
tgccactacc aacgatatga tcggt                                           25


<210>  11
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  11
ccgcttcacc ttgaattgct tctac                                           25


<210>  12
<211>  19
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  12
ccccctgaag cttcaccgg                                                  19


<210>  13
<211>  21
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  13
cctacttgcg ctgcatgtgc c                                               21


<210>  14
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  14
ccagcggcgg ggtcaggtgc gttac                                           25


<210>  15
<211>  27
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  15
gagccggcga aggtccagga atgtttg                                         27


<210>  16
<211>  26
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  16
ggtgctgccg gtgatcgagg agttga                                          26


<210>  17
<211>  26
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  17
cgccgcagac gcacccgagg agacac                                          26


<210>  18
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  18
ccagcggcgg ggtcaggtgc gttac                                           25


<210>  19
<211>  24
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  19
ccagaggccg ggcgtgtgat actt                                            24


<210>  20
<211>  27
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  20
aacctcattc aatttgggca agcatag                                         27


<210>  21
<211>  32
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  21
accactctac ttccgtcgaa tataatactg gc                                   32


<210>  22
<211>  29
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  22
gctgactttg tatcacggga gacctagat                                       29


<210>  23
<211>  32
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  23
ttcttataat gtgtttgcat aatgccatcg ta                                   32


<210>  24
<211>  27
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  24
tcaccacttt cccaagacac ggctaca                                         27


<210>  25
<211>  32
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  25
gctgctagta caggctctcg caatgactac ac                                   32


<210>  26
<211>  32
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  26
ggatctttag gaacgctata acgccattag ga                                   32


<210>  27
<211>  32
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  27
cactgtgctg gagaagtaga tagaacgcat tt                                   32


<210>  28
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  28
gctgtctact tgggagtgat ttgag                                           25


<210>  29
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  29
ggacccaaag cccacaacct aaata                                           25


<210>  30
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  30
cttggtcagt tgggtggata gtagt                                           25


<210>  31
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  31
gtggcagggt ctatgatgga actaa                                           25


<210>  32
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  32
gggtcctagt gatggtatct gaaca                                           25


<210>  33
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  33
gagggaactt agaggatggg tcaat                                           25


<210>  34
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  34
ttgggaggat aggtgacaga agcat                                           25


<210>  35
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  35
tagggaaaga gcaagagcgg acaat                                           25


<210>  36
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  36
ctccatatcc caccctaaga acaac                                           25


<210>  37
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  37
cgctcaactt cccattacat cctac                                           25


<210>  38
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  38
ttcccagctc tgctttgtgt agttg                                           25


<210>  39
<211>  25
<212>  DNA
<213>  ARTIFICIAL SEQUENCE

<220>
<223>  SYNTHETIC OLIGONUCLEOTIDE

<400>  39
gccttcctct aactctgcct tcata                                           25


